Fig. 1

mTORC1 upregulates the expression of ERO1α. A Venn diagram analysis of differentially upregulated genes in four datasets. Dataset 1: Tsc2 − / − vs. Tsc2 + / + MEFs. Dataset 2: Tsc1 − / − vs. Tsc1 + / + MEFs. Dataset 3: ELT3 cells treated with DMSO vs. ELT3 cells treated with rapamycin (20 nM, 24 h). Dataset 4: hiPSCs Tsc2 − / − vs. hiPSCs Tsc2 + / + . B Tsc2 + / + and Tsc2 − / − MEFs were treated with rapamycin (Rapa, 20 nM) or DMSO for 24 h. C Tsc1 + / + and Tsc1 − / − MEFs were treated with rapamycin (Rapa, 20 nM) or DMSO for 24 h. B and C Cell lysates were subjected to immunoblotting with the indicated antibodies (left panels); ERO1α levels were analyzed by qRT–PCR (right panels). D IF analysis of the expression of ERO1α in the indicated cells. Scale bar, 20 μm. E and F Tsc2 − / − (E) or Tsc1 − / − (F) MEFs were transfected with siRNA targeting mTOR (simTOR), Raptor (siRaptor) or the control (siNC) for 48 h. G Tsc2 + / + or Tsc1 + / + MEFs were treated with 5 μM MHY1485 for 24 h. E–G Cell lysates were subjected to immunoblotting with the indicated antibodies. H A representative kidney of Tsc2 + / − mice. Red arrows indicate renal cysts and cystadenomas. I Representative IHC images of p-S6 and ERO1α staining from renal cystadenomas of Tsc2 + / − mice. Error bars indicate mean ± SD of triplicate samples. ****P < 0.0001